Compositions and methods related to antibodies to staphylococcal protein a

a staphylococcal protein and antibody technology, applied in the field of immunology, microbiology, pathology, can solve the problem that the fda approved staphylococcal disease vaccine is currently unavailable, and achieve the effect of reducing activity

Active Publication Date: 2014-06-19
UNIVERSITY OF CHICAGO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0079]As used herein, the term “modulate” or “modulation” encompasses the meanings of the words “inhibit.”“Modulation” of activity is a decrease in activity. As used herein, the term “modulator...

Problems solved by technology

An FDA approved vaccine that prevents st...

Method used

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  • Compositions and methods related to antibodies to staphylococcal protein a
  • Compositions and methods related to antibodies to staphylococcal protein a
  • Compositions and methods related to antibodies to staphylococcal protein a

Examples

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example 1

[0256]Monoclonal Antibodies to Staphylococcus Aureus Protein A

[0257]SpAKKAA-mAbs Protect Mice Against Staphylococcal Disease.

[0258]BALB / c mice were immunized with purified SpAKKAA using a prime-booster regimen and antigen specific IgG responses were quantified by ELISA. Animals were euthanized and their splenocytes fused with myeloma cells. The resulting hybridomas were screened for the production of antigen-specific mAbs. Initially, protein A-specific mAbs were screened using the functional assays as well as the murine infection model (Table 1). After the initial screen, we selected three mAbs (5A10, 3F6, and 3D11) for further characterization as these antibodies displayed the best immune protection in each isotype group (Table 1). BALB / c Mice were immunized with affinity purified mAbs (5 mg·kg−1 body weight) and challenged by injecting 1×107 CFU S. aureus Newman, a methicillin-sensitive clinical isolate (MSSA) (Baba et al., 2007), into the periorbital venous sinus of the right eye...

example 2

Materials and Methods

[0299]Bacterial Strains and Growth Conditions.

[0300]S. aureus strains Newman and MW2 were grown in tryptic soy broth (TSB) at 37° C. Escherichia coli strains DH5a and BL21 (DE3) were grown in Luria-Bertani (LB) broth with 100 μg·ml-1 ampicillin at 37° C.

[0301]Monoclonal Antibodies.

[0302]Mouse monoclonal antibodies were generated by the conventional method (Köhler, G., and C. Milstein. 1975). Briefly, BALB / c mice (8 week old, female, Jackson Laboratory) were immunized by intraperitoneal injection with 100 μg purified SpAKKAA emulsified 1:1 with Complete Freund's Adjuvant (CFA, DIFCO). On days 21 and 42, mice were boosted by intraperitoneal injection with 100 μg of the same antigen emulsified 1:1 with Incomplete Freund's Adjuvant (IFA, DIFCO). On days 31 and 52, mice were bled and serum samples screened by ELISA for specific antibodies. Seventy-nine days following initial immunization, mice that demonstrated strong antigen-immunoreactivity by ELISA were boosted wi...

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Abstract

Embodiments concern methods and compositions for treating or preventing a bacterial infection, particularly infection by a Staphylococcus bacterium. Aspects include methods and compositions for providing a passive immune response against the bacteria. In certain embodiments, the methods and compositions involve an antibody that binds Staphylococcal protein A (SpA).

Description

[0001]This application claims priority to U.S. Provisional Patent Application Ser. No. 61 / 523,751 filed on Aug. 15, 2011, Ser. No. 61 / 615,083 filed on Mar. 23, 2012, Ser. No. 61 / 618,417 filed on Mar. 30, 2012, and Ser. No. 61 / 674,135 filed on Jul. 20, 2012, all of which are incorporated herein by reference in their entirety.[0002]This invention was made with government support under AI52747 and AI92711 from the National Institute of Allergy and Infectious Diseases (NIAID) and 1-U54-AI-057153 awarded by the National Institutes of Health. The government has certain rights in the invention.FIELD OF THE INVENTION[0003]The present invention relates generally to the fields of immunology, microbiology, and pathology. More particularly, it concerns methods and compositions involving antibodies to bacterial proteins and bacterial peptides used to elicit such antibodies. The proteins include Staphylococcal protein A (SpA).BACKGROUND[0004]The number of both community acquired and hospital acqu...

Claims

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Application Information

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IPC IPC(8): C07K16/42A61K45/06A61J1/00A61K39/40
CPCC07K16/4233A61K45/06A61K39/40A61J1/00A61K38/00A61K2039/505A61K2039/507A61P31/00A61P31/04A61P37/00C07K16/1271C07K2317/76C07K2317/92A61K39/395A61K2300/00A61K39/085A61K2039/545C07K2317/14C07K2317/21C07K2317/51C07K2317/515C07K2317/56C07K2317/565C07K2317/569G01N33/6854G01N2333/31G01N2500/04
Inventor SCHNEEWIND, OLAFMISSIAKAS, DOMINIQUE M.KIM, HWAN KEUNEMOLO, CARTADEDENT, ANDREA
Owner UNIVERSITY OF CHICAGO
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